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A new model to assess analgesic activity: Pain‐

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 DRUG DEVELOPMENT RESEARCH 28:169-175 (1993)

Research Article

A New Model to Assess Analgesic Activity: Pain-Induced

Functional Impairment in the Rat (PIFIR)
Francisco J.Lopez-Munoz, Luis A. Salazar, Gilberto Castaneda-Hernandez, and
Julian E. Villarreal
Seccidn de Terapeutica Experimental, Departamento de Farmacologia y Toxicologia, Centro
de lnvestigacidn y de Estudios Avanzados del lnstituto Politecnico Nacional, Mexico
(L.A.S., C.C.-H., I.E.V.);and Departamento de Sistemas Bioldgicos, Universidad
Autdnoma Metropolitana-Xochimilco, Mexico (F.).L.-M.)

ABSTRACT A new experimental model to assess analgesic activity of both analgesic and non-
steroidal antiinflammatory drugs is described. It uses the unilateral intra-articular knee injection of
an uric acid suspension in mineral oil to produce acute inflammation, pain, and functional motor
impairment. The model, named "pain-induced functional impairment in the rat" (PIFIR) assesses
analgesic activity by measuring the capacity to walk with the injured extremity. The procedure
determines both the potencies of analgesic drugs and the time course of the effect. Analgesia of
selected reference agents was followed for 4 h and the effect versus time curves were constructed.
The area under the curve (effect versus time), an expression of the overall activity during the
observation period, increased in a dose-dependent manner. The area under the curve, Em,,, T,,,,,
and ED,, of reference agents tested are reported. The PlFlR procedure was sensitive to opiate and
nonopiate analgetics (nonsteroidal antiinflammatory drugs) and possibly steroidal antiinflammatory
drugs. These characteristics make it suitable for screening purposes. o 1993 wiley-~iss,I ~ C .

Key Words: pain model, analgesia, PIFIR, opioids, nonsteroidal antiinflammatory drugs, rat

INTRODUCTION in nociception during tonic pain states is required if

Understanding of the mechanisms of analgesia
and t h e development of novel analgesic agents de-
pends to a major extent on the available animal meth-
ods for t h e quantitative determination of pain. The
majority of t h e traditional assays for analgesic activity
measure transient rather than continuous pain. These
models vary widely in the type of noxious stimuli ap-
plied. Some of these use thermal stimuli [D'Amour
and Smith, 1941; Woolfe and McDonald, 19441, while
others use mechanical [Bianchi and Franceschini,
1954; Collier e t al., 1961; Randall and Selitto, 19571,
electrical [Turker and Turker, 1970; Ayhan e t al.,
19831. and chemical stimulation [Siegmund e t al..
T h e relevance of phasic endpoints yielded by
acute stimuli to clinical pain has been questioned, as
the in the aforementioned methods ar' ar-
tificial noxious stimuli [Pircio e t al., 19751. A better
understanding of the physiological processes involved
novel analgesics are to b e properly evaluated
[Wheeler-Aceto and Cowan, 19911.
There are several models of persistent pain. O n e
is adjuvant-induced polyarthritis in the rat [Pearson,
19561. This procedure has been well characterized
[ D e Castro Costa e t al., 19811 and has proven to b e
useful for the assay of analgesic agents [Pircio e t al.,
19751. However, the induction of polyarthritis results
in an extended systemic involvement and therefore
observed responses cannot b e confidently considered
as d u e to nociception alone [Wheeler-Aceto and
Cowan, 19911.
Received final version October 29, 1992; accepted Novem-
ber 2, 1992,
Address reprint requests to Francisco 1. L6pez-MuAoz, De-
partamento de Sistemas Biol6gicos, Universidad Aut6norna Met-
ropo~itana-Xoch~mi~co, Apartado Postal 23-181 col, Sari juan,
Delegation Xochimilco 16000 Mexico D.F., Mexico.

63 1993 Wiley-Liss, Inc.

 Alternative methods have been described, such
as intra-articular or subcutaneous injections of a vari-
ety of substances in a hind limb [Faires and McCarty,
1962; Pardo and Rodriguez, 1966; Van Arman et al.,
1970; Okuda et al., 1984; Wheeler-Aceto and Cowan,
19911. These procedures have been useful for both
the assay of analgesic drugs and the characterization of
the inflammatory process.
To adequately evaluate the potential efficacy of
an analgesic agent, it is necessary to determine not
only the analgesic efficacy at a fixed time but the du-
ration of the effect [Mattok, 19881. Methods that allow
the determination of the time course of analgesia will
facilitate characterization of the mechanisms of action
of analgesic drugs and pharmacokinetic/pharmacody-
namic studies.
Pardo and Rodriguez [I9661 developed a method
in dog which allowed them to quantitatively follow
the analgesic action with time. Another procedure de-
veloped by Van Arman et al. [I9701 allowed a quan-
titative estimation of analgesia, but all measurements
were taken at fixed time. Okuda et al. [I9841 reported
a modified version of this procedure using cats that
allowed them to follow the time course of morphine in
a quantitatively fashion. These three procedures, al-
though they are suitable for the determination of an-
algesia over time, have the disadvantage that animals
must be submitted to lengthy training periods to ob-
tain reliable results. Nonetheless these procedures
are useful in studying the physiological bases of pain
and analgesia but are impractical for screening pur-
poses.
In the present study, we report a method of
pain-induced functional impairment in the rat
(PIFIR). This procedure determines the potency and
duration of analgesic action produced by various
classes of compounds by measuring their effect on
inflammation-induced tonic pain and permits the
quantitative evaluation of the kinetics of the analgesic
effect.
MATERIALS AND METHODS
Animals
Female Wistar rats, 180-220 g, from our own
breeding facilities [Crl: (WI)BR] were used in this
study. Twelve hours before experiments, food was
withheld with free access to water. All procedures
performed followed the recommendations of The
Committee for Research and Ethical Issues of the In-
ternational Association for the Study of Pain [1980].
Measurement of Analgesic Activity
The animals were anesthetized with ether in an
anesthesia chamber (glass dryer Pirex saturated with
ether vapors). Pain was induced by an intra-articular
injection of 0.05 ml of 30% uric acid suspended in
mineral oil in the knee joint of the right hind limb.
The suspension was prepared by grinding 3.0 g of uric
acid with 10 ml of mineral oil in a glass mortar and
pestle (Pirex). The intra-articular injection was per-
formed through the patellar ligament using a 1 ml
glass syringe (Becton, Dickinson LTDA, Brazil) with a
24 gauge needle of 5 mm. Immediately, an electrode
was attached to each hind paw between the plantar
pads. Rats were allowed to recover from anaesthesia
and were then placed on a stainless steel cylinder of
30 cm diameter. The cylinder was rotated at 4 RPM,
forcing the rats to walk. Training periods were not
necessary because the rats learning in the first min-
utes. The variable measured was the time of contact
between each of the rat's hind paws and the cylinder.
When the electrode placed on the animal's paw made
contact with the cylinder floor, a circuit was closed
and the time that the circuit remained closed was
recorded. The cylinder was rotated for 2 min periods,
during which time recordings were made with 30 min
rest periods between recordings. In all subsequent
experiments, analgesic agents were administered 2.5
h after uric acid injection. Thus, this time was con-
sidered as time zero for measurements of analgesic
effect. Drugs were given at this time and the time of
contact was measured every 30 min for 4 h. All ex-
periments were performed between 7:00 am and
14:OO. The animals were then sacrificed.
Instrumentation
The rotary walking device consisted of a hollow
cylinder made of stainless steel of 30 cm diameter and
60 cm long. The cylinder was divided into six individ-
ual tracks with annular pieces of stainless steel of 48
cm of external diameter. Each track was covered with
stainless steel wire mesh (20 mesh per inch) to pro-
vide a nonslip surface for the animals. This arrange-
ment allowed the study of six animals at the time. An
electrical motor (Westinghouse Elec. Corp. Model
309P384-A) with electronic control rotated the cylin-
der at a constant speed of 4 RPM.
The time of contact of each hind paw was re-
corded by plantar electrodes. Each electrode con-
sisted of a drop of silver solder in a disk-like form
placed at the end of a 26 gauge insulated copper wire.
The metallic disk was glued to a piece of rubber film
that was adhered to the hind paw with cyano-acrylic
adhesive to prevent direct contact between the metal
and the skin. The other end of the copper wire was
connected to the time counting device. The schematic
diagram used for counting the time of contact be-
tween the electrodes and the cylinder floor is shown
 ASSAY FOR ANALGESIC ACTIVITY: PlFlR

Figure 1. Schematic diagram of the system used for recording of the time of contact between the
rat's hind paws and the cylinder floor. A: Time counting device, B: switch box, C: switches, D:
electrodes, E: rotary walking stainless steel cylinder, F: rotating motor drive.

in Figure 1. An Apple@ I1+ computer equipped with
a Mountain@ AID board and a switch box was used to
collect data by means of a Basic software routine (Fig.
2).
Drugs
Uric acid was purchased from Sigma Chemical
Co. (St. Louis, MO), Dipyrone sodium (Hoechst),
morphine hydrochloride (Mexican Secretariat of
Health), pentazocine hydrochloride (Winthrop Prod-
ucts), d-propoxyphene hydrochloride (ElinLilly), and
hydrocortisone 21-phosphate (Sigma Chemical Co.,
St. Louis, MO) were dissolved in distilled water. Ace-
tylsalicylic acid (Miles) and acetaminophen (Sigma
Chemical Co., St. Louis, MO) were suspended in
0.5% carboxymethyl cellulose. Indomethacin (Merck,
Sharp and Dohme) was dissolved in a sodium bicar-
bonate solution. Adequate controls were performed
with each of the used vehicles. Doses of each agent
are referred to the particular salt.
Data Analysis
Data are expressed as the functionality index
(FI). This is the time of contact of the injected limb
divided by the time of contact of the control left limb
multiplied by 100.
The maximal observed effect (Em,,) can be ex-
pressed in terms of FI and the time required to reach
this response (T,,,,), as shown in Table 1. These pa-
rameters reflect the potency and the rate of onset of
drug action by a given route. Furthermore, since the
time course of the effect has been followed, it is pos-
sible to use the cumulative analgesic effect during the
whole observation period as the area under the curve
(AUC) in a similar manner to that used in pharmacok-
inetics to express the amount of drug absorbed
[Gibaldi and Perrier, 19751.
Since the AUC value represents the integrated
analgesic effect during the observation period and
thus includes both the maximal response and the du-
ration of action, this expression was preferred for the
construction of dose-response curves. The maximal
possible AUC value that could be achieved under the
experimental conditions was 375 area units (% . h).
This is due to the fact that the maximal FI is 100% and
the observation period consisted of 4 h with the first
determination being performed at 0.5 h. Considering
that the FI reached 100% in 0.5 h and remained at
this value until the hour 4, the AUC obtained is 375
units (obtained with the trapezoidal rule and taking in
account the area of the first triangle).
All values in the text and figures are mean of at
least six animals + SE. The area under the curve
172 LOPEZ-MUNOZ ET AL.

PROGRAM IN BASIC TO GATHER DATA IN THIS MODEL pathological analysis showed edema and inflamma-
10 TEXT: HOME: CLEAR: PRINT "<<<PIFIR>>>":PRINT tory infiltration in synovial membranes with no effect
20 INPUT "NAME OF DATA FILE? (RETURN = NONE) ":NO$
30 IF NOS = "" THEN 60 on the joint cartilage (data not shown). Groups of
40 N = 451: INPUT "NUMBER OF READINGS? (451 = 2 MINUTES) ": R$
50 IF R$ < > "" THEN N = VAL (R$) polymorphonuclear leukocytes forming masses or free
60 DIM D(121.V(l21.C(12)
70 REM *;**'SET UP.ADDRESS VECTOR +*+* filaments were present in the articular cavity. Fibrin,
80 FOR I = 1 TO 12
90 D(1) = 49315 + I:V(I) = 0: NEXT hemorrhage, or necrosis were absent and no synovial
100 C = Dfl): FOR I = 1 TO 11:D(I) = D(I+l): NEXT:D(lZ) = C
110 REM **** CLEAN UP READING ***I cell proliferation was detected. The intensity of the
120 FOR I = 1 TO 1 2 : ~= PEEK (~(1)): NEXT inflammatory lesion was moderate as reflected by for-
130 IF NO$ = "" THEN 420
140 REM ++++ READING LOOP 1 * * * * 1 * * 1 * * * * " ' * "
150 FOR I = 1 TO N: FOR J = 1 TO 12
mation of polymorphonuclear leukocyte exudate.
- - - C = PEEK ID(JI\
160 ~ - ,~ - ,
170 IF C-2-175 THEN V(J) = V(J) + 1
180 IF C > 175 THEN C(J) = C(J) + 1 Time Course of Functionality lndex (FI)
190 NEXT : NEXT : PRINT CHRS(7)
200
-~ REM- - **** DISPLAY
~- VALUES O N SCREEN ********
~
The time course of FI was followed for 6.5 h
210 HOME
220 VTAB 1: PRINT "CONTROL INJECTED" measuring every 30 min (Fig. 3). FI was 100% at the
230 FOR J = 2 TO 12 STEP 2: VTAB J: PRINT V(J), V(J -1): NEXT
240 IF NOS = "" THEN END start of the experiments, i.e., the time of contact (sec)
250 INPUT "SAVE DATA? (Y/N) ":R$: IF R$ = "N" THEN 340
260 PRINT CHRS(4) "OPEN "NO$: ",D2" between both hind limbs was similar. Control rats
270 PRINT CHRS(4) "WRITE "NO$
280 PRINT CHRS(41 "CLOSE "NOS injected with 0.05 ml of mineral oil did not show any
290 PRINT C H R S ~ "APPEND
~ ~ "NOS
300 PRINT c H ~ S ij4 "WRITE "NOS ' significant decrease (P > 0.05) of FI during the whole
310 FOR I = 1 TO 12
320 PRINT V(1): NEXT period of observation. Intra-articular injection of uric
330 PRINT CHRS(~)"CLOSE "NOS
340 PRINT: PRINT; INPUT "ANOTHER READING? (Y/N) W : RS acid suspension induced a dysfunction of the injected
350 IF R$ = "N" THEN 392
360 FOR I = 1 TO 12 limb that was apparent as a gradual reduction of FI
370 V(I) = O:C(I) = 0: NEXT
~ - --- ~~. -~ -
with a maximal decrease at 2.5 h leveling at zero for
390
392
GOT0 120
PRINT: PRINT : INPUT "ANOTHER TEST READING? (Y/N)";R$
the remainder of experiment (6.5 h). Morphine (10
393 IF RS = "N" THEN 400 mg/kg sc) administered at 2.5 h when FI approached
394 FOR I = 1 TO 12:V(I) = O:C(II. . = 0: NEXT
395
396
HOME: CLEAR :NO$ =
GOT0 30
zero produced a gradual recovery. Values reached a
400
410
PRINT CHRS(4 ) "RUN DIRECTORY ,Dl"
REM **** SYSTEM TEST ****
maximum of about 75% and then decreased slowly.
420 C5 = 175: HOME: PRINT CHRS(12)
430
440
VTAB 1: PRINT "CONTROL INJECTED"
N = 30: FOR I = 1 T O N: FOR J = 1 TO 12
Effect of Morphine Administered by Various Routes
450 C = PEEK (D(J))
460 IF C < C5 THEN VfJl = V(J1 + 1 The FI values obtained with morphine after i.p.,
470 IF c > c5 THEN c i ~ j= c i ~ j+ 1
480 NEXT i.m., and p.o. administrations are depicted in Figure
490 FOR J = 2 TO 12 STEP 2: VTAB J: PRINT V(J),V(J-1): NEXT
500 NEXT : PRINT CHRS(7): CHRS(7): GOT0 340 4. Doses of 10 mg/kg i.m. or 31.6 mgkg i.p. yielded
results similar to those observed with 177.8 mg/kg
Figure 2. Program in basic t o gather data in t h e PIFIR. p.o. (P > 0.05). FI reached a maximum of about 70%
to 80% in 1 h and then decreased gradually. An oral
dose of 177.8 mg/kg resulted in a relatively rapid in-
(AUC) relating FI versus time was obtained by the crease of FI over a 1.5 h period, followed by a sus-
trapezoidal rule [Gibaldi and Perrier, 1975; Rowland tained effect for the remainder of experiment. These
and Tozer, 19891. Control rats and rats injected with results were consistant with the pharmacokinetic pro-
uric acidlmineral oil were compared by analysis of file of morphine [Jaffe and Martin, 19911.
variance (ANOVA) and Dunnett's test. The AUCs of
morphine administered by differents routes were Effect of Reference Agents on the Functionality lndex
compared using ANOVA. Values of P < 0.05 were The time-action course of selected analgesic and
considered statistically significant. ED,, values were antiinflammatory agents on the FI are shown in Fig-
calculated by a polynominal program fitting of two ure 5. Morphine (5.6 mg/kg sc) induced a rapid in-
variables in iterative form taking in account the max- crease in FI which reached a maximum of about 70%
imal effect possible in this model, 375 units of AUC. in 1 h and then decreased. Acetylsalicylic acid (316.2
The E,,, and T,,, were obtained directly from the mdkg p.0.) produced a maximal functionality index of
FI against time curves of doses that produced the 45% in 1 h and values remained stable for 4 h. Ace-
highest AUC of each drug. taminophen (177.8 mg/kg p.0.) increased the FI by
27% in 1 h followed by a rapid decline to zero in 3 h.
Indomethacin (2.4 mg/kg sc) induced a slow but
RESULTS
steady increasing FI, reaching 70% in 4 h. For this
Histopathology reason the maximal effect of indomethacin could not
Intra-articular injection of uric acid in the right be established within the 4 h observation period. Hy-
hind knee produced local acute inflammation. Histo- drocortisone (50 mdkg sc) exhibited a latency period
 ASSAY FOR ANALGESIC ACTIVITY: PlFlR 173

TABLE 1. Some Pharmacological Parameters of Opiates and Nonsteroidal Antiinflammatory Drugs Determined in the PlFlR Model
Drug and ED, in
dose ErnaXa T~rnaxa AUC PI FIR^
(mglkg) (FI) X 2 SE (h) (% . h) X & SE (mg/kg) X 2 SE
Morphine ( 1 7.8 sc) 96.6 12.6 1.5 310.2 25.3 7.9 1.3
Pentazocine (31.6 sc) 78.2 26.5 3 .O 241.8 72.1 30.2 5.4
d-Propoxyphene (31 6.2 sc) 87.2 6.0 4.0 204.1 17.2 208.2 4.1
lndomethacin (3.16 sc) 74.1 10.9 3 .O 227.4 30.1 2.6 1.3
ASAC (562.3 p.0.) 83.9 7.5 2.5 258.3 54.6 399.6 1.9
Dipyrone (1,000 sc) 87.0 5.2 2.5 268.9 15.34 517.5 1.2
"Variables measured of curves of time course.
b~arameterobtained taking as Em, the general maximal effect in this model in the present form (375 area units).
'Acetylsalicylic acid.

oy' I

0 1 2 3 4
0 'Morphins 4 6 T lM E (hours)
T lM E (hours)
Figure 4. Time course of FI in rats injected with uric acid treated
Figure 3. Time course of FI. (O), Control rats (intra-articular in- by morphine 2.5 h after uric acid injection. (0)10 mglkg i.m.; (0)
jection of 0.05 ml of mineral oil in the right hind knee).,).( Uric 31.6 mglkg i.p.; (W) 177.8 mglkg p.0. Symbols represent the mean
acid rats (intra-articular uric acid in the right hind knee). (n),
Uric & SE for at least eight animals. P > 0.05 between these groups.

acid-morphine rats (10 mglkg of morphine sc 2.5 h after uric acid

injection). Data are presented as mean 2 SE of six animals. P >
0.05 between control rats and temporal course of rats injected
with mineral oil.
of 1.5 h, prior to any effect being seen, and reached a
maximum of 40% in 3-4 h. The doses used were se-
lected from dose-response curves obtained previ-
ously.
The observations with different compounds sug-
gest that the temporal patterns of drug action may
reflect the pharmacological properties characteristic
to a class of compounds. For example analgesic agents
had a fast onset while antiinflammatory compounds
were slower acting.
Expression and Analysis of Experimental Data
The results with morphine, pentazocine, d-
propoxyphene, indomethacin, and dipyrone were ad-
ministered sc and acetylsalicylic acid given orally are
presented in Figure 6. In all cases AUC increased in
a dose-dependent manner. Indomethacin and mor-
phine appeared to be the most potent of the assayed
compounds but indomethacin showed a limited effi-
cacy with a maximum at 227 area units. Pentazocine
yielded a parallel curve but shifted to the right com-
pared to that of morphine. A lower potency as well as
a limited efficacy were observed with d-propoxy-
phene, which reached its maximum at about 200
area units. Acetylsalicylic acid p.o. and dipyrone sc
yielded similar dose-response curves. They were con-
siderably less potent than indomethacin but they ex-
hibited a higher efficacy. A summary of parameters as
such Em,,, T,,,,, AUC, and the ED,, of the drugs
from the data shown in Figure 6 are documented in
Table 1.
DISCUSSION
The PIFIR method appears to be suitable for the
assay of analgesic drugs but is not suitable to charac-
terize agents such as indomethacin because of their
delayed onset and long duration of action. The deter-
174 LOPEZ-~ufioz
ET AL.

were similar to those reported with sodium urate-

induced effects in dogs [Van Arman et al., 19701 and
cats [Okuda et al., 19841.
It has been reported that the leg dysfunction is
mainly produced by the inflammatory process be-
cause it was reversible by antiinflammatory agents
[Van Arman et al., 19701. However, Okuda et al.
[I9841 have provided evidence using morphine that
animals avoid using the traumatized extremity chiefly
because of a manifestation of pain. In our experiments
we observed that drugs without antiinflammatory ac-
tions, such as morphine, exhibited high potency, and
0 1 2 3 4 efficacy. Conversely, pure antiinflammatory agents,
T I M E (hours) such as hydrocortisone, showed a different activity
and the onset of the effect was very slow (indometh-
Figure 5. Time course of FI in rats injected with uric acid treated
acin was active but was observed over too short a
by analgesic and antiinflammatory agents. ( a ) Morphine 5.6 mg/
kg sc; (0) acetylsalicylic acid 316.2 mg/kg p.0.; (H) acetami- period). These results are in agreement with those of
nophen 177.8 mglkg p.0.; (0) indomethacin 2.4 mglkg sc, and Okuda et al. [I9841 and suggest that leg dysfunction is
( A ) hydrocortisone sc 50 mglkg. Symbols represent the mean 2 primarily due to pain caused by acute inflammation.
SE for at least six animals. Uric acid injection produced a persistent dys-
function of the injured limb which lasted at least for 4
h (Fig. 3). Therefore, the procedure can be consid-
ered as a model of tonic pain and thus may simulate
clinical pain in temporal parameters.
The use of the FI has the advantage that the
noninjured limb serves as control. Drugs having ef-
fects on the motor activity of the animals should also
affect the functionality of the control limb; therefore
the use of the F I enables to minimize this type of
interference. The FI is determined when rats are
forced to walk. This is a natural behavior of the ani-
mals and thus lengthy training sessions are not nec-
essary. Rats get used to the rotating apparatus in less
than 2 h. The use of small animals as experimental
subjects presents the advantage that significant
Figure 6. Dose-response curves of several analgesic agents de- amounts of drug are not required.
termined in the PIFIR. The response is expressed as AUC (vertical Many pharmacological assays, like those for
axis) estimated over a 4 h observation period. Left panel (A): ( a )
morphine sc, (B)pentazocine sc, and ( A ) d-propoxyphene sc.
drugs affecting the cardiovascular, respiratory, gas-
Right panel (B): (0) indomethacin sc, (A)dipyrone sc, and (0) trointestinal, or endocrine systems [Holford and
acetylsalicylic acid p.0. Symbols correspond to the mean 2 SE for Sheiner, 19811, permit the evaluation of efficacy as a
at least six determinations. function of time. In the area of analgesia, traditional
procedures, such as the "tail flick" or the "tail clip,"
evaluate behavioral responses like squeaking or biting
mination of the time course of study would accommo- after application of an acute stimulus. Therefore, after
date such compounds. several stimuli there is a conditioned learning effect
Histopathological analysis showed that uric acid interfering with the response. In the PIFIR method,
induced acute inflammatory response comparable to the noxious stimulus is applied only once and pro-
that seen with intra-articular injection of sodium urate duces a constant pain level.
crystals [Faires and McCarty, 1962; Van Arman et al., In addition to variations in potency, there were
19701. After uric acid injection, there was a motor important differences in the time courses of the anti-
dysfunction of the injected extremity, i.e., the rats nociceptive effect induced by the agents studies.
avoided the use of the injured hind limb when forced Morphine (ip) had a fast onset which diminished after
to walk. The onset of dysfunction was gradual with a reaching a maximum. Acetylsalicylic acid (po) re-
maximal (almost 100%) effect in 2.5 h. These features sulted in a more persistent effect, although the doses
 ASSAY FOR ANALGESIC ACTIVITY: PlFlR 175

needed were quite high. Indomethacin (sc) induced D'Amour FE, Smith DL (1941): A method for determining loss of
an effect of considerable magnitude but of slow onset. pain sensation. J Pharmacol Exp Ther 72:74-79.
From the data in Figure 5, it is misleading to De Castro Costa MC, De Sutter P, Gybels J, Van Haes J (1981):
consider the analgesic efficacy at a fixed arbitrary time Adjuvant-induced arthritis in rats: A possible animal model for
chronic pain. Pain 10:173-185.
(for example 2 h after drug administration). The con-
struction of FI versus time curves allows the direct Faires JS, McCarty DJ (1962):Acute arthritis in man and dog after
intrasynovial injection of sodium urate crystals. Lancet ii:682-
determination of the maximal effect as well of the time 684.
to reach the maximum (Table 1).
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The AUC is a parameter that considers the over- Dekker, Inc., pp 293-296.
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the duration of the effect. For this, some drugs can relationship: Clinical application of pharmacokinetic-pharmaco-
have the Em, similar (i.e., d-propoxyphene and dipy- dynamic models. Clin Pharmacokinet 6:429-453.
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